Technical Field
The present disclosure relates to a heterologous DNA barcoding method, and more specifically to a heterologous DNA barcoding method for analyzing large amounts of gene samples in a simultaneous and parallel manner.
Background Art
The advent of next-generation DNA sequencing technologies has led to a dramatic increase in the number of sequences that can be read at one time. DNA barcoding based on these next-generation technologies aims at parallel reading of a great number of DNA sequences at one time. For example, Matthias Meyer and colleagues reported different methods for simultaneous analysis of two or more samples by attaching DNA barcodes to the samples by ligation (Matthias Meyer et al., Targeted high-throughput sequencing of tagged nucleic acid samples, Nucleic Acids Research 35, e97 (2007), Matthias Meyer et al., Parallel tagged sequencing on the 454 platform, Nature Protocols 3, 267-278 (2008)). Ligation of each barcode possessing an inherent sequence onto a target DNA allows for the insertion of additional information into the target DNA. Accordingly, when a large number of DNAs are analyzed, the information inherent to the barcode-tagged DNAs can also be read. For example, barcoding can be used to determine from what cells specific DNAs are extracted.
The separation and parallel analysis of different DNAs requires barcoding of the DNAs. Traditional barcoding methods necessitate the operations of constructing different DNA oligos having inherent barcode sequences and ligating the oligos to DNAs. However, traditional oligo synthesis methods entail considerable construction costs of various DNA oligos. Further, the oligos should be sequentially ligated to DNAs. This operation requires much time and labor. Large amounts of materials are also needed for the reactions, incurring considerable costs. Thus, improvements are needed.